Comparing seminal plasma biomarkers between normospermic and azoospermic men

Azoospermia affects more than 10% - 15% of infertile male subjects attending infertilty clinics. At present, testicular biopsy is the golden standard procedure for evaluating spermatogenesis status in men with azoospermia. Semen collection and analysis is a non-invasive method and has proven to be valuable in the evaluation of spermatogenesis. Identification of seminal plasma markers with testicular or extra-testicular origins have a great value in predicting the prescence of sperm in testicular tissue and presumptive cause of azoospermia. The aim of this study was to find such markers by comparing the content of seminal plasma using different methods in normospermic and azoospermic men. Semen samples were collected from 200 men attending Avicenna Infertility Clinic [AIC] in Tehran, Iran. Semen samples were analysed according to WHO guidlines. The subjects were divided into two groups: normospermic [n = 100; group one] and azoospermic men [n = 100; group two] according to semen analysis results. Seminal plasma was separated by high speed centrifuagation and stored in -20°C. Four markers including fructose, neutral alpha glucosidase [NaG], inhibin B and anti-Mullerian hormone [AMH] were measured in seminal plasma. Fructose and NaG were evaluated by spectrophotometry, while inhibin B and AMH were assessed by ELISA method. The spermatogenesis status in the azoospermic group was evaluated by histopathological method following testicular biopsy. Fructose concentration showed no difference between the two groups. However, it was significantly correlated with sperm count [p < 0.01, r = -0.408]. Seminal plasma inhibin B [OR: 1.01; 95%: CI: 1.005 - 1.016], AMH [OR: 1.63; 95% CI: 1.17 - 2.28] and N alpha G, [OR: 1.07; 95% CI: 1.04 - 1.1] levels were higher in normospermic subjects compared to azoospermic men. There were significant differences in inhibin B and AMH concentrations between the two groups based on the presence or absence of mature sperm in testicular biopsies [p < 0.01]. Inhibin B concentration was positively correlated with sperm count in the normospermic group, however, N alpha G concentration correlated with sperm count of normospermic men [p < 0.01, r = 0.345] and the subjects'age in both groups. Inhibin B and AMH were correlated with the presence of sperm in testicular tissue samples. According to non-specific changes in inhibin B and AMH concentrations, identification of more specific molecular markers in seminal plasma to definitely evaluate the status of spermatogenesis is recommended

Effects of sperm chromatin integrity on fertilization rate and embryo quality following intracytoplasmic sperm injection

Sperm chromatin integrity has been being recognized as an important factor in male fertility. During normal fertilization, high quality sperm with intact chromatin are selected through natural selection in journey from vagina to fallopian tube. However, using Assisted Reproductive Techniques, particularly ICSI, the natural selection is bypassed. Therefore sperm with DMA breakage have the opportunity to fertilize the egg which may lead to decreased embryo quality and implantation rate. The aim of this study was to evaluate the effects of sperm chromatin integrity on ICSI outcomes. A total of 200 semen samples were collected from couples undergoing ICSI and were analyzed according to WHO criteria. Each sample was evaluated for sperm chromatin integrity using four cytochemical assays and semen processing by swim up method. The ICSI was carried out according to a long-term pituitary down-regulation protocol. The correlation between sperm parameters, sperm chromatin integrity and ICSI outcomes [fertilization rate and embryo quality] was examined. The mean number of oocyte, fertilization rate and cleavage embryos per cycles was 7.5 +/- 5.0, 74.O6% +/- 25 and 5.4 +/- 3.6, respectively. There was not significant correlation between the results of chromatin assays [AO, AB, TB, and CMA3] and fertilization outcomes following ICSI. The fertilization rate was significantly higher for a group with less than 10% chromatin abnormality [p<0.05]. Sperm chromatin integrity is essential for successful fertilization, embryo development and normal pregnancy. A protamine deficiency appeared to affect fertilization rate and embryo quality. However, the presence of confounding factors such as selection of spermatozoa according to normal morphology may influence the effect of sperm chromatin status on ICSI outcomes